Mosaic Drosophila

The point of mosaicisms techniques is to manipulate gene function at the cellular level without disturbing other cellular subpopulations. In Drosophila, genetic mosaicisms are generated by mitotic recombination. Flp-FRT derived systems are widely used due to its high specific and recombination rates. If you have a specific gene locus, we recommend Flp-out recombination, where mutations are not restricted by cell cycle and gene locus. Relying on our powerful strategy platform, we provide you with flexible, visual, inducible, and site-specific mosaicisms, including dominant and cryptic markers. We devote into adapting to clients' needs of mosaic analysis in various environments and conditions.

• Flp-FRT Derived Systems

While there are many recombinases and integrases that induce targeted recombination in vivo. We mainly use Flp-FRT recombination, the main tool for creating mosaics in Drosophila. Flp induces recombination between FRT sites, both in cis and trans, depending on the relative positions of the 2 FRT sites. A key requirement for mosaic analysis is the ability to detect mutant cells with non-lethal and easily trackable cellular markers. We combine Gal4-UAS as well as Gal80 to build the Mosaic Analysis with a Repressible Cell Marker (MARCM) system that flexibly label mutant cell lines to meet customers' requirements for real-time, high-resolution tracking analysis. We offer a wide range of easy-to-track and sensitive labels, such as fluorescent protein, CD2, etc.

Technical Characteristics

• Limitations on the fourth chromosome or for any genes located between the FRT and the centromere.
• Visualization with real-time tracking.
• High accuracy, high specificity, and short project cycle.

• Flp-out Derived Systems

Flp-out recombination is a type of cis-recombination that occurs on the same chromosome, cutting DNA fragments between FRTs, independent of gene site and time constraints. The excision event is heritable. The earlier it occurs, the greater the number of mutant cells and the greater the extent of the effect. We offer a variety of Flp-out variant techniques, such as insertion of STOP frames between FRTs, separation of ubi promoters from target genes or markers (G-TRACE), and Mutant Analysis by Rescue Gene Excision (MARGE) to research any gene in Drosophila genome in principle, etc. There are no limitations of our projects. Just provide your gene name and research purpose, we will solve your mosaic fly construction problem and help your research.

Technical Characteristics

• Not limited by gene site and time.
• Suitable for the research of relatively complex gene that can not be studied by gene knock-out
• High accuracy, high specificity, and short project cycle.
• Enables invisible and explicit flexible tagging.

The genetic mosaic and pedigree tracking system is flexible and changeable. According to your specific genetic characteristics and research needs, we pursue an integrated access strategy to generated more powerful, sensitive, multi-sites, and visually traceable, mosaic research models, in the way of combining and improving Flp strategies under spatial- and temporal- control.

• MOSAICISMS STRATEGY

We provide you with the most convenient and effective strategy for your research purpose. Full communication and years of experience allow us to make a comprehensive plan, detailing selection of optimal FRT sites, optimizing Flippase codon sequences, adding selectable marker, assembly of appropriate Flp-FRT composed system, production of stable transgenic fruit flies to maximize the chance of success. Moreover, we provide other one-stop services for further gene function or interaction researches.

• CONSTRUCTS

Depending on the project approach, we offer optimal transgenic vectors generation and purification. We provide common detectable markers used in mosaic analysis (yellow, CD2, fluorescent proteins, β-galactosidase, etc.). FRT or FRT variant sites are produced by precise modification of endogenous or transgenic genomic technology. Unless specifically requested, we inject wild-type fruit flies.

• VALIDATION

As standard, we validate at the genomic level with advanced sequencing and we provide a functional readout that verifies system efficiency and flies' vitality. We first validate whether the gene recombination occurs at the design site, further ensure the genotype of the derived homozygous cell, and the connection of all genetic elements at the far end of the FRT site, to achieve reliable cell-autonomous labeling and cloning. And we confirm whether the marker is expressed or whether the ubiquitous reporter gene expression is twice as high as the background.

• PHENOTYPIC ANALYSIS 

We make sure all mosaic models work well and achieve the purpose of the experiment. The effect of mosaicism is evaluated using various techniques to quantify mRNA or from the protein level (or by omics) due to clients' need. Furthermore, analysis can also be determined by visualizing protein immunofluorescence (optimal).

We provide regular updates and in-time messages if any issue or complication happen during the manufacturing process. The final report details every manufacturing process and analysis data will feed back to you with the flies.

We ship final transformants in either larvae or adults. The final transformants vials are maintained in the facility for two weeks as backup.

At CD BioSciences, we offer complete services to improve and extend clients' researches in Drosophila. We pioneer advanced approaches to assist customers around the world access high-quality solutions and supports.

• According to your specific genetic characteristics and research needs, we offer mosaic model customization which is more powerful, sensitive, multi-sites, and visually traceable.
• We provide complex overlapping mosaicisms construction and analysis services, which help to study the development of complex tissues.
• By constructing a series of vectors carrying different markers and ubiquitous promoters, we provide the strategy of real-time tracking phenomena of multiple genes labels in whole body tissues or cells.
• Further connection services, including sample preparation, immunofluorescence imaging analysis and other qualitative and quantitative phenotypic analysis services.

• Division of cell lineages and division of cellular autonomic and involuntary gene
• The interaction of different cell groups
• Developmental Biology